Effect of neurokinin 3 receptor agonist senktide on the activation of hypothalamic nNOS neurons in cycling female rats

Abstract

Objective: To examine the effect of an neurokinin 3 receptor (NK3R) agonist, senktide, on neuronal nitric oxide synthase (nNOS) activation in the median eminence-arcuate nucleus (ME-ARC) and preoptic area (POA) regions of the hypothalamus across proestrus, diestrus, and ovariectomized states in female rats and its correlation with luteinizing hormone (LH) secretion.

Methods: Adult female Sprague-Dawley rats were examined for proestrus and diestrus phases of the estrous cycle. Female rats were categorized into proestrus and diestrus groups, and each was further divided into four subgroups (n=4). In both the diestrus and proestrus categories, Group 1 was the control group. Groups 2, 3, and 4 received senktide (100 μg/kg^-1), NK3R antagonist SB222200 (10 mg/kg^-1), and SB222200 followed by senktide, respectively. To evaluate the effect of sex steroids on NK3R agonist-induced nNOS activation, female rats underwent bilateral ovariectomy and were divided into four groups (n=3). Group 1 served as the control. Group 2 received a subcutaneous injection of 17 β-estradiol 3-benzoate (E2, 3 μg/rat). Group 3 received E2 and progesterone (30 μg/rat). Group 4 was administered senktide (100 μg/kg). Female rats from each group were sacrificed, blood was collected for LH ELISA, and hypothalamic tissues were collected for Western blotting.

Results: Senktide increased nNOS phosphorylation in the ME-ARC during both the proestrus and diestrus phases. In the POA, senktide increased nNOS phosphorylation only during the diestrus phase. In ovariectomized rats, senktide activated nNOS independent of sex steroid levels. Senktide also increased serum LH concentration in diestrus and ovariectomized female rats.

Conclusions: Senktide, an NK3R agonist, activates nNOS in the POA and ME-ARC regions of the hypothalamus in a phase dependent manner. The activation of nNOS by senktide suggests a potential mechanism by which neurokinin B triggers nNOS activation in the ARC and POA regions and regulates GnRH/LH secretion.

Keywords

Neurokinin B / Neuronal nitric oxide synthase / Luteinizing hormone / Neurokinin 3 receptor / Ovariectomized / Preoptic area / Median eminence-arcuate nucleus

Conflict of interest statement

The authors have no relevant financial or non-financial interests to disclose.

Acknowledgment

JP is thankful to the University Grants Commission (UGC) for the start-up grant F.30-318/2016 (GP-105) and the Department of Science (DST)-Science and Engineering Research Board (SERB) for the Early Carrier Research Grant ECR/2015/000240 (GP-90) and Core Research Grant CRG/2020/003257 (GP-176). VP thanks DST-SERB for the Junior and Senior Research Fellowship. TA and RS also thank CSIR for the JRF and SRF for conducting this research. VP would also like to sincerely thank the Central University of Punjab, notably the Central Instrument Facility and Animal House Facility, for providing the essential infrastructure and state-of-the-art facilities that have enabled me to conduct my research. JP is also grateful to the Central University of Punjab for RSM-CUPB/CC/16/00/13 for providing financial assistance, space, and infrastructure to carry out this research.

Funding

This work is supported by DST-Science and Engineering Research Board (SERB) Early carrier research grant ECR/2015/000240, Core research grant CRG/2020/003257, the University Grants Commission (UGC start-up grant F.30-318/2016), and the Central University of Punjab RSM grant-CUPB/CC/16/00/13.

Data availability

The data of the current study is available from the corresponding author on a reasonable request.

Authors’ contributions

Jyoti Parkash and Vikash Prashar contributed to the conception and design. Vikash Prashar conducted material preparation, experiments, data collection, and analysis. The original manuscript draft was written by Vikash Prashar and reviewed by all authors. Tania Arora and Randeep Singh assisted with sample collection and draft writing. Arti Sharma assisted in data analysis, writing, supervision, and review of the manuscript. Jyoti Parkash contributed resources, provided supervision, and reviewed and approved the final manuscript.

Publisher’s Note

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