DOI: 10.5061/dryad.rn8pk0pmz
title: Data from: Selective depletion of kisspeptin neurons in the hypothalamic arcuate nucleus in early juvenile life reduces pubertal LH secretion and delays puberty onset in mice

description: Puberty is the critical developmental transition to reproductive
 capability driven by the activation of gonadotropin‐releasing hormone
 (GnRH) neurons. The complex neural mechanisms underlying pubertal
 activation of GnRH secretion still remain unknown, yet likely include
 kisspeptin neurons. However, kisspeptin neurons reside in several
 hypothalamic areas and the specific kisspeptin population timing pubertal
 onset remains undetermined. To investigate this, we strategically
 capitalized on the differential ontological expression of
 the Kiss1 gene in different hypothalamic nuclei to
 selectively ablate just arcuate kisspeptin neurons (aka KNDy neurons)
 during the early juvenile period, well before puberty, while sparing RP3V
 kisspeptin neurons. Both male and female transgenic mice with a majority
 of their KNDy neurons ablated (KNDyABL) by diphtheria toxin treatment in
 juvenile life demonstrated significantly delayed puberty onset and lower
 peripubertal LH secretion than controls. In adulthood,
 KNDyABL mice demonstrated normal in vivo LH pulse
 frequency with lower basal and peak LH levels, suggesting that only a
 small subset of KNDy neurons is sufficient for normal GnRH pulse timing
 but more KNDy cells are needed to secrete normal LH concentrations. Unlike
 prior KNDy ablation studies in rats, there was no alteration in the
 occurrence or magnitude of estradiol‐induced LH surges in KNDyABL female
 mice, indicating that a complete KNDy neuronal population is not essential
 for normal LH surge generation. This study teases apart the contributions
 of different kisspeptin neural populations to the control of puberty
 onset, demonstrating that a majority of KNDy neurons in the arcuate
 nucleus are necessary for the proper timing of puberty in both sexes.
description: # Data from: Selective depletion of kisspeptin neurons in the hypothalamic
 arcuate nucleus in early juvenile life reduces pubertal LH secretion and
 delays puberty onset in mice
 [https://doi.org/10.5061/dryad.rn8pk0pmz](https://doi.org/10.5061/dryad.rn8pk0pmz) ## Description of the data and file structure **General information:** 1.      **Title of Dataset:** “Data from: Selective depletion of kisspeptin neurons in the hypothalamic arcuate nucleus in early juvenile life reduces pubertal LH secretion and delays puberty onset in mice” 2.      **Contact Author Information:** Dr. Alexander S. Kauffman, Department of OBGYN and Reproductive Sciences, University of California San Diego, 9500 Gilman Drive, #0674, La Jolla, California 92093. email: [akauffman@ucsd.edu](mailto:akauffman@ucsd.edu) 3.      **Corresponding Publication:** The manuscript containing this data has been peer-reviewed and accepted for publication: “Eulalia A Coutinho, Lourdes A Esparza, Paige H Steffen, Reanna Liaw, Shreyana Bolleddu, Alexander S Kauffman. Selective depletion of kisspeptin neurons in the hypothalamic arcuate nucleus in early juvenile life reduces pubertal LH secretion and delays puberty onset in mice. The FASEB Journal, Oct 15;38(19):e70078. doi: 10.1096/fj.202401696R **Description of the data and file structure:** The data are from a study that compared reproductive hormone levels, puberty onset, body weights, and kisspeptin neuron counts between several groups of male and female transgenic mice that were treated in juvenile life with diphtheria toxin to selectively ablate their arcuate nucleus KNDy neuron population or control mice, as described in detail in the Methods section of the published manuscript (Coutinho et al., “Selective depletion of kisspeptin neurons in the hypothalamic arcuate nucleus in early juvenile life reduces pubertal LH secretion and delays puberty onset in mice, The FASEB Journal, 2024”). ### Files and variables #### File: All\_figures\_data\_for\_KissiDTR\_study.xlsx **Data File Description:**  The dataset used to generate all graphs in the figures for the associated manuscript are deposited here in an .xlsx file "All_figures_data_for_KissiDTR_study.xlsx". The .xlsx file comprises 11 separate worksheets named with the corresponding manuscript Figure number (e.g., Fig 2A, Fig 3, Fig 4A, etc.) which indicates which specific figure or figure panels the data was used for graphs in the published manuscript, along with a short identifying text in the worksheet name (e.g. KNDy neurons, female puberty, adult LH, body weights, etc.) which gives additional context about what the data in that worksheet corresponds to. Missing values are denoted as blank cells. All methods and analyses are described in detail in the published manuscript Methods section. ##### Abbreviations *  tdtom, tdtomato (red) fluorescence in kisspeptin cells * ARC, arcuate nucleus of the hypothalamus * PND, postnatal day (age after birth) * LH, luteinizing hormone * BW, body weight * g, grams * ng/ml, nanograms per milliliter * KNDy, Kisspeptin/NKB/Dynorphin-expressing neurons * RP3V, rostral periventricular nucleus of the 3rd ventricle, a region in the hypothalamus * VO, vaginal opening (day of; an early stage pubertal marker) * FE, first estrus (day of; a late stage pubertal marker) * PPS, preputial separation (day of; a pubertal marker in males) * DT, diphtheria toxin (used to ablate kisspeptin neurons) * AM, morning period * PM, evening period * Cre-, mice lacking Cre recombinase expression in kisspeptin neurons * Cre+, mice expressing Cre recombinase in kisspeptin neurons * KNDyABL, mice with KNDy neurons ablated * CON, control mice **Treatment Groups:** * control, control mice with no ablation of KNDy neurons * KNDyABL, experimental mice with KNDy neurons ablated by DT treatment in juvenile life **Variables (and units):** **Figure 1C**: Fluorescent tdtomato-expressing cells (number of). These data are the number of tdtomato-expressing cells detected in transgenic Kiss-tdtom mice in the ARC or RP3V brain regions at either PND 12 or PND 28 ages. **Figure 2C, F, I**: Fluorescent Kiss1 mRNA-expressing cells (number of). These data are the numbers of fluorescent Kiss1 mRNA-expressing cells detected in either control or KNDyABL mice in the ARC or RP3V brain regions in adult males (Fig 2C) or adult females (Fig 2F, 2I). **Figure 3A-F**: Body weight (g). These data are the body weights (BW; measured in g) of either control or KNDyABL male and female mice at different postnatal ages. Female BWs are graphed in Figs 3A, B, C) and males BW graphed in (Figs 3D, E, F).   **Figure 4A-F**: Vaginal Opening (day of), First Estrus (day of), Body weight (g). These data are pubertal measures in female control and KNDyABL mice. VO (day of) is graphed in Fig 4B, BW on the day of VO (g) is graphed in Fig 4C, and FE (day of) is graphed in Figure 4E. **Figure 5A-C:** Preputial Separation (day of), Body weight (g). These data are pubertal measures in male control and KNDyABL mice. PPS (day of) is graphed in Fig 5B and BW on the day of PPS (g) is graphed in Fig 5C. **Figure 6A, B**: Luteinizing hormone (LH) levels in the blood (ng/ml blood) These data are the blood levels of LH (measured in ng LH/ml blood) of either peri-pubertal control or KNDyABL female (Fig 6A) and male (Fig 6B) mice at postnatal day (PND) 28. **Figure 7A-D**: Luteinizing hormone (LH) levels in the blood (ng/ml blood) These data are the blood levels of LH (measured in ng LH/ml blood) of either control or KNDyABL mice in adulthood. LH levels of adult females at either baseline (Fig 6A) or after Kisspeptin injection (post-KP; Fig 6B). LH levels of adult males at either baseline (Fig 6C) or after Kisspeptin injection (post-KP; Fig 6D). **Figure 8A-F**: Pulsatile secretion parameters of Luteinizing hormone (LH) secretion These data are for different parameters of pulsatile secretion patterns of LH of control or KNDyABL male mice in adulthood. Figure 8A depicts blood LH levels (ng/ml) over time (minutes) of 2 control males or 2 KNDyABL males. Fig 8B is mean LH levels (ng/ml) across the entire 66 minutes of sampling. Fig 8C is LH Pulse frequency (# of LH pulse events per hour); Fig 8D is basal LH levels (ng/ml), Fig 8E is peak LH levels during a pulse (ng/ml) and Fig 8F is pulse amplitude (ng/ml), which is the difference between pulse peak and basal. **Figure 9A, B**: Luteinizing hormone surge (LH) secretion (ng LH/ml blood) These data are LH surge secretion of control or KNDyABL female mice. Figure 9A depicts blood LH levels (ng/ml) in control or KNDyABL females at either the AM or PM time-points. Fig 8B is the % of females in each treatment group demonstrating PM LH levels above the LH surge threshold of 0.7 ng/ml.