Browsing by Author "Kazuyoshi Tsutsui"

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Age-dependent variation in the RFRP-3 neurons is inversely correlated with gonadal activity of mice
(Academic Press Inc., 2010) Sumit Sethi; Kazuyoshi Tsutsui; Chandra Mohini Chaturvedi
The present study analyzed changes in the expression of RFamide-related peptide-3 (RFRP-3; a mammalian ortholog of avian gonadotropin-inhibitory hormone), in the brain and correlated it with testicular activity of mice of different age groups (day-old, 1-, 3-, 5-, 7-, 9-, 11-, 13-week and 1.5-year-old). Testicular activity after a progressive increase up to 13-week of age declined in the old mice. On the other hand, while immunoreactive (ir) RFRP-3 neurons were not seen in the day-old mice, few appeared in 1-week-old mice, their number and size increased drastically at 3-week of age. This condition remained unaltered until 7-week of age followed by a progressive decline up to the age of 13-week and thereafter increased again in the old age. The present findings indicate that hyperactivity of the ir-RFRP-3 neurons of dorsomedial nucleus of hypothalamus (DMH) observed in prepubertal mice declines in reproductively active mice and increases again in the old mice having declined reproductive performance. It is concluded that aging mice exhibits inverse correlation of RFRP-3 neurons and gonadal activity suggesting that function of RFRP-3 is not initiated until 1-week of age and thereafter it could participate in the regulation of gonadal development. © 2010 Elsevier Inc.
Apoptosis-mediated testicular alteration in Japanese quail (Coturnix coturnix japonica) in response to temporal phase relation of serotonergic and dopaminergic oscillations
(Company of Biologists Ltd, 2016) Somanshu Banerjee; Kazuyoshi Tsutsui; Chandra Mohini Chaturvedi
Reproductive performance of many avian species, including Japanese quail, is reported to be modulated by specific temporal phase relation of serotonergic and dopaminergic oscillations. Accordingly, it has been shown that the serotonin precursor 5-HTP and the dopamine precursor L-DOPA given 8 h apart induce gonadal suppression and given 12 h apart lead to gonadal stimulation, while other temporal relationships were found to be ineffective. In the present study, we investigated the effects of 8- and 12-h phase relation of neural oscillations on testicular responses including expression of GnRH-I, GnIH, pro-apoptotic proteins (p53 and Bax), inactive and active executioner caspase-3, and the uncleaved DNA repair enzyme PARP-1. Testicular volume and mass decreased significantly in 8-h quail and increased in 12-h quail compared with controls. Expression of ir-GnIH, p53, Bax and active-caspase-3 increased and that of GnRH-I, pro-caspase-3 and uncleaved PARP-1 decreased in 8-h quail compared with controls. A TUNEL assay also confirmed testicular regression in these quail. Testes of 12-h quail exhibited significantly increased expression of GnRH-I, pro-caspase- 3 and uncleaved PARP-1 compared with the control group. Our findings suggest that differential response of avian testes to 8- and 12-h phase relation of serotonergic and dopaminergic neural oscillations may be attributed to autocrine/paracrine action of GnIH expression, which is upregulated in regressed testes, leading to apoptotic changes, and downregulated in developed testes, causing apoptotic inhibition. It is concluded that specific phase relation of neural oscillations may modulate the local testicular GnRH-GnIH system and alter the apoptotic mechanism in quail testes. Moreover, these findings highlight the physiological effects of time-dependent drug delivery, including the specific time intervals between two drugs. © 2016. Published by The Company of Biologists Ltd.
Central and peripheral neuropeptide RFRP-3: A bridge linking reproduction, nutrition, and stress response
(Academic Press Inc., 2022) Padmasana Singh; Shabana Anjum; Raj Kamal Srivastava; Kazuyoshi Tsutsui; Amitabh Krishna
This article is an amalgamation of the current status of RFRP-3 (GnIH) in reproduction and its association with the nutrition and stress-mediated changes in the reproductive activities. GnIH has been demonstrated in the hypothalamus of all the vertebrates studied so far and is a well-known inhibitor of GnRH mediated reproduction. The RFRP-3 neurons interact with the other hypothalamic neurons and the hormonal signals from peripheral organs for coordinating the nutritional, stress, and environmental associated changes to regulate reproduction. RFRP-3 has also been shown to regulate puberty, reproductive cyclicity and senescence depending upon the nutritional status. A favourable nutritional status and the environmental cues which are permissive for the successful breeding and pregnancy outcome keep RFRP-3 level low, whereas unfavourable nutritional status and stressful conditions increase the expression of RFRP-3 which impairs the reproduction. Still our knowledge about RFRP-3 is incomplete regarding its therapeutic application for nutritional or stress-related reproductive disorders. © 2022 Elsevier Inc.
Changes in GnRH I, bradykinin and their receptors and GnIH in the ovary of Calotes versicolor during reproductive cycle
(Academic Press Inc., 2008) Padmasana Singh; Amitabh Krishna; Rajagopala Sridaran; Kazuyoshi Tsutsui
The aim of this study was to investigate changes in the abundance of gonadotrophin releasing hormone I (GnRH I) and GnRH I receptor in the ovary of Calotes versicolor during the reproductive cycle and correlate them with the changes in gonadotrophin inhibitory hormone (GnIH), bradykinin and bradykinin B2 receptor in order to understand their interaction during ovarian cycle. GnRH I, bradykinin and their receptors and GnIH, were localized immunohistochemically in the ovary. Relative intensity of these peptides was estimated from the contralateral ovary using slot/Western blot followed by densitometry. The immunostaining of GnRH I, bradykinin and their receptors and GnIH were localized in the granulosa cells of previtellogenic follicles and stroma cells, whereas in the peripheral part of the cytoplasm in oocytes of vitellogenic and ovulatory follicles. The GnRH I immunostaining was relatively higher in inactive phase, but was low during active preovulatory phase suggesting inverse correlation with circulating estradiol level. The study showed a positive correlation between the expression pattern of GnRH I and GnIH, but showed a negative correlation between GnIH with GnRH I receptor in the ovary. This study further suggests a possibility for bradykinin regulating GnRH I synthesis in the ovary. An increase in the immunostaining of both GnRH I and GnIH in the oocyte prior to ovulation suggests their involvement in the oocyte maturation. It is thus concluded that the ovary of C. versicolor possesses GnRH I-GnIH-bradykinin system and interaction between these neuropeptides may be involved in the regulation of follicular development and oocyte maturation. © 2008 Elsevier Inc.
Direct effects of RFRP-1, a mammalian GnIH ortholog, on ovarian activities of the cyclic mouse
(Academic Press Inc., 2017) Anushree Dave; Amitabh Krishna; Kazuyoshi Tsutsui
Arg(R)-Phe(F)-amide related peptide-1 (RFRP-1) and -3 (RFRP-3) are known as mammalian orthologs of gonadotropin-inhibitory hormone (GnIH). In mammals, these RFRPs are expressed not only in the hypothalamus and but also in gonads. Inhibitory roles of the hypothalamic and gonadal RFRP-3 in reproduction have been documented in mammals. However, functional roles of the hypothalamic and gonadal RFRP-1 in reproduction are still unclear in mammals. Therefore, in vitro studies were conducted to elucidate the direct effect of RFRP-1, a mammalian GnIH ortholog, on ovarian activities, such as steroidogenesis, apoptosis, cell proliferation and metabolism in the cyclic mouse. The ovaries collected from the proestrus mice were cultured in vitro with different doses (Control, 1 ng/ml, 10 ng/ml and 100 ng/ml) of RFRP-1 for 24 h at 37 °C. A significant dose-dependent increase in estradiol release from the ovary was detected after the treatment of RFRP-1. Therefore, changes in the ovarian activities, such as steroidogenic markers (luteinizing hormone receptors; LH-R and 3β-hydroxysteroid dehydrogenase; 3β-HSD), apoptotic markers [Poly(ADP-ribose) polymerase-1; PARP-1 and cysteine-aspartic protease; caspase-3], a cell proliferation marker (proliferating cell nuclear antigen; PCNA) and metabolic markers (GLUT-4; glucose uptake) were assessed by the treatment of RFRP-1 in the proestrus ovary. The densitometry analysis showed the treatment of RFRP-1 significantly increased the expressions of LH-R and 3β-HSD, steroidogenic markers. In contrast, the expressions of PCNA, a cell proliferation maker; PARP-1 and caspase-3, apoptotic markers were significantly decreased. Interestingly, RFRP-1 treatment further increases significantly glucose uptake and GLUT-4 receptor expression. These findings indicate that RFRP-1 possesses a stimulatory effect on ovarian steroidogenesis in the proestrus mouse. This is the first evidence showing the direct action of RFRP-1 on steroidogenesis in any vertebrate. In addition, RFRP-1 may also act directly on ovarian folliculogenesis as an inhibitory factor. © 2017 Elsevier Inc.
Effects of gonadotropin-inhibitory hormone on folliculogenesis and steroidogenesis of cyclic mice
(2011) Padmasana Singh; Amitabh Krishna; Kazuyoshi Tsutsui
Objective: To evaluate the effects of gonadotropin-inhibitory hormone (GnIH) treatment on ovarian activity of mice. Design: Animal study. Setting: Reproductive physiology laboratory of university department of zoology. Animal(s): Twelve-week-old female mice of inbred Parkes strain. Intervention(s): Mice treated with different doses of GnIH (control, 100 ng, 500 ng, and 2 μg per day) for 8 days were studied. For in vitro study, the ovaries of proestrus mice were cultured with different doses of GnIH for 24 hours at 37°C. Main Outcome Measure(s): Folliculogenesis, steroidogenesis, luteogenesis, and apoptosis in the ovaries of control and GnIH-treated mice. Result(s): GnIH treatment produced significant changes in body mass, circulating steroid levels, and ovarian activity in the mice. GnIH also caused dose-dependent histologic changes in follicular development and luteinization. The antral follicles showed abnormal changes. The mice treated with increasing dose of GnIH showed significant changes in steroid synthesis owing to inhibitory effects of GnIH on ovarian expression of LH receptor, steroidogenic acute regulatory, and 3β-hydroxysteroid dehydrogenase proteins. Conclusion(s): GnIH inhibited follicular development and steroidogenesis in the ovary of mice. This study thus suggests biologic significance of this neuropeptide in regulating ovarian activity. © 2011 American Society for Reproductive Medicine, Published by Elsevier Inc.
Immunohistochemical localization of GnRH and RFamide-related peptide-3 in the ovaries of mice during the estrous cycle
(2011) Padmasana Singh; Amitabh Krishna; Rajagopala Sridaran; Kazuyoshi Tsutsui
Gonadotropin releasing hormone (GnRH) has now been suggested as an important intraovarian regulatory factor. Gonadotropin inhibitory hormone (GnIH) a hypothalamic dodecapeptide, acts opposite to GnRH. GnRH, GnIH and their receptors have been demonstrated in the gonads. In order to find out the physiological significance of these neuropeptides in the ovary, we aim to investigate changes in the abundance of GnRH I and GnIH in the ovary of mice during estrous cycle. The present study investigated the changes in GnRH I, GnRH I-receptor and RFRP-3 protein expression in the ovary of mice during estrous cycle by immunohistochemistry and immunoblot analysis. The immunoreactivity of GnRH I and its receptor and RFRP-3 were mainly localized in the granulosa cells of the healthy and antral follicles during proestrus and estrus and in the luteal cells during diestrus 1 and 2 phases. The relative abundance of immunoreactivity of GnRH I, GnRH I-receptor and RFRP-3 undergo significant variation during proestrus and thus may be responsible for selection of follicle for growth and atresia. A significant increase in the concentration of RFRP-3 during late diestrus 2 coincided with the decline in corpus luteum activity and initiation of follicular growth and selection. In general, immunolocalization of GnRH I, GnRH I-receptor and RFRP-3 were found in close vicinity suggesting functional interaction between these peptides. It is thus, hypothesized that interaction between GnRH I-RFRP-3 neuropeptides may be involved in the regulation of follicular development and atresia. © 2011 Springer Science+Business Media B.V.
Inhibitory roles of the mammalian GnIH ortholog RFRP3 in testicular activities in adult mice
(BioScientifica Ltd., 2014) Shabana Anjum; Amitabh Krishna; Kazuyoshi Tsutsui
The aim of this study was to evaluate the effects of in vivo and in vitro treatments with RFamide-related peptide 3 (RFRP3), a mammalian gonadotropin-inhibitory hormone ortholog, on testicular activities, i.e. spermatogenesis and steroidogenesis, in mice. Mice were treated in vivo with different doses of RFRP3 (control: 0.02 μg, 0.2 μg, and 2.0 μg/day) for 8 days. For in vitro study, the testes of mice were evaluated with different doses of RFRP3 (control: 1 and 10 ng/ml) with or without LH (control: 10 and 100 ng/ml) for 24 h at 37 °C. RFRP3 treatment produced significant changes in the body mass, circulating steroid level, and testicular activity in mice. RFRP3 treatment also caused dose-dependent histological changes in spermatogenesis, such as decline in germ cell proliferation and survival markers and increase in apoptotic markers in testis. Both in vivo and in vitro studies showed the inhibitory effect of RFRP3 on testosterone synthesis in the testis. RFRP3 inhibited the expression of the receptor for LH (LHCGR), STAR protein, cytochrome P450 side-chain cleavage (CYP11A1) and 3ß-hydroxysteroid dehydrogenase in the testis, and testosterone secretion dose dependently. This study also suggested that the inhibitory effect of RFRP3 in the testis may be mediated through local production of GnRH. Thus, RFRP3 inhibits testicular steroidogenesis and spermatogenesis either indirectly through GnRH or by directly influencing germ cell proliferation, survival, and apoptosis. © 2014 Society for Endocrinology.
Localization of Gonadotropin-Releasing Hormone (GnRH), Gonadotropin-Inhibitory Hormone (GnIH), Kisspeptin and GnRH Receptor and Their Possible Roles in Testicular Activities From Birth to Senescence in Mice
(2012) Shabana Anjum; Amitabh Krishna; Rajagopala Sridaran; Kazuyoshi Tsutsui
The changes in distribution and concentration of neuropeptides, gonadotropin-releasing hormone (GnRH), gonadotropin-inhibitory hormone (GnIH), kisspeptin, and gonadotropin-releasing hormone receptor (GnRH-R) were evaluated and compared with reproductive parameters, such as cytochrome P450 side-chain cleavage (P450 SCC) enzyme activity, androgen receptors (AR) in the testis and serum testosterone levels, from birth to senescence in mice. The results showed the localization of these molecules mainly in the interstitial and germ cells as well as showed significant variations in immunostatining from birth to senescence. It was found that increased staining of testicular GnRH-R coincided with increased steroidogenic activity during pubertal and adult stages, whereas decreased staining coincides with decreased steroidogenic activity during senescence. Similar changes in immunostaining were confirmed by Western/slot blot analysis. Thus, these results suggest a putative role of GnRH during testicular pubertal development and senescence. Treatment with a GnRH agonist ([DTrp6, Pro9-NEt] GnRH) to mice from prepubertal to pubertal period showed a significant increase in steroidogenic activity of the mouse testis and provided further support to the role of GnRH in testicular pubertal maturation. The significant decline in GnRH-R during senescence may be due to a significant increase in GnIH synthesis during senescence causing the decrease in GnRH-R expression. It is considered that significant changes in the levels of GnRH-R may be responsible for changes in steroidogenesis that causes either pubertal activation or senescence in testis of mice. Furthermore, changes in the levels of GnRH-R may be modulated by interactions among GnRH, GnIH, and kisspeptin in the testis. © 2012 Wiley Periodicals, Inc.
Possible role of GnIH as a mediator between adiposity and impaired testicular function
(Frontiers Media S.A., 2016) Shabana Anjum; Amitabh Krishna; Kazuyoshi Tsutsui
The aim of the present study was to evaluate the roles of gonadotropin-inhibitory hormone (GnIH) as an endocrine link between increasing adiposity and impaired testicular function in mice. To achieve this, the effect of GnIH on changes in nutrients uptake and hormonal synthesis/action in the adipose tissue and testis was investigated simultaneously by in vivo study and separately by in vitro study. Mice were treated in vivo with different doses of GnIH for 8 days. In the in vitro study, adipose tissue and testes of mice were cultured with different doses of GnIH with or without insulin or LH for 24 h at 37°C. The GnIH treatment in vivo showed increased food intake, upregulation of glucose transporter 4 (GLUT4), and increased uptake of triglycerides (TGs) in the adipose tissue. These changes may be responsible for increased accumulation of fat in white adipose tissue, resulting in increase in the body mass. Contrary to the adipose tissue, treatment with GnIH both in vivo and in vitro showed decreased uptake of glucose by downregulation of glucose transporter 8 (GLUT8) expressions in the testis, which in turn resulted in the decreased synthesis of testosterone. The GnIH treatment in vivo also showed the decreased expression of insulin receptor protein in the testis, which may also be responsible for the decreased testicular activity in the mice. These findings thus suggest that GnIH increases the uptake of glucose and TGs in the adipose tissue, resulting in increased accumulation of fat, whereas simultaneously in the testis, GnIH suppressed the GLUT8-mediated glucose uptake, which in turn may be responsible for decreased testosterone synthesis. This study thus demonstrates GnIH as mediator of increasing adiposity and impaired testicular function in mice. © 2016 Anjum, Krishna and Tsutsui.
RF-amide related peptide-3 (RFRP-3): a novel neuroendocrine regulator of energy homeostasis, metabolism, and reproduction
(Springer Science and Business Media B.V., 2021) Shabana Anjum; Muhammad Nasir Khan Khattak; Kazuyoshi Tsutsui; Amitabh Krishna
A hypothalamic neuropeptide, RF-amide related peptide-3 (RFRP-3), the mammalian ortholog of the avian gonadotropin-inhibitory hormone (GnIH) has inhibitory signals for reproductive axis via G-protein coupled receptor 147 in mammals. Moreover, RFRP-3 has orexigenic action but the mechanism involved in energy homeostasis and glucose metabolism is not yet known. Though, the RFRP-3 modulates orexigenic action in co-operation with other neuropeptides, which regulates metabolic cues in the hypothalamus. Administration of GnIH/RFRP-3 suppresses plasma luteinizing hormone, at the same time stimulates feeding behavior in birds and mammals. Likewise, in the metabolically deficient conditions, its expression is up-regulated suggests that RFRP-3 contributes to the integration of energy balance and reproduction. However, in many other metabolic conditions like induced diabetes and high-fat diet obesity, etc. its role is still not clear while, RFRP-3 induces the glucose homeostasis by adipocytes is reported. The physiological role of RFRP-3 in metabolic homeostasis and the metabolic effects of RFRP-3 signaling in pharmacological studies need a detailed discussion. Further studies are required to find out whether RFRP-3 is associated with restricted neuroendocrine function observed in type II diabetes mellitus, aging, or sub-fertility. In this context, the current review is focused on the role of RFRP-3 in the above-mentioned mechanisms. Studies from search engines including PubMed, Google Scholar, and science.gov are included after following set inclusion/exclusion criteria. As a developing field few mechanisms are still inconclusive, however, based on the available information RFRP-3 seems to be a putative tool in future treatment strategies towards metabolic disease. © 2021, The Author(s), under exclusive licence to Springer Nature B.V. part of Springer Nature.
Temporal phase relation of circadian neural oscillations alters rfamide-related peptide-3 and testicular function in the mouse
(2010) Sumit Sethi; Kazuyoshi Tsutsui; Chandra Mohini Chaturvedi
In order to study the effect of the temporal synergism of neural oscillations on reproductive regulation and the response of RFamide-related peptide-3 (RFRP-3; a mammalian ortholog of avian gonadotropin-inhibitory hormone), expression of immunoreactive RFRP-3 in the neurons of the dorsomedial nucleus of the hypothalamus was monitored in sexually immature and mature laboratory mice (study I). In study II, the effects of serotonin and dopamine precursors (5-hydroxytryptophan and L-dihydroxyphenylalanine; injected daily, 8 or 12 h apart, for 13 days in 3-week-old mice) on testicular activity and immunoreactive RFRP-3 neurons were studied until 24 days after treatment. Results indicate high levels of expression of immunoreactive RFRP-3 in the sexually immature and 8-hour mice (simulating gonadal suppression), while a low level was noted in mature and 12-hour mice (simulating gonadal stimulation). These findings not only suggest the modulation of gonadal development in mice (during the course of puberty attainment) by changing the temporal phase relation of serotonergic and dopaminergic oscillations (as in some seasonally breeding species), but also demonstrate an inverse correlation of RFRP-3 neurons and gonadal activity in both control and experimental conditions. Copyright © 2009 S. Karger AG.