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The tissue distribution of nucleobindin-2/nesfatin-1 in the reproductive organs of bitches with regard to the animal’s age and body weight Cover

The tissue distribution of nucleobindin-2/nesfatin-1 in the reproductive organs of bitches with regard to the animal’s age and body weight

Journal of Veterinary Research
Volume 69 (2025): Issue 1 (March 2025)
By:
Open Access
|Mar 2025

Figures & Tables

Fig. 1.

Relative expression of Nucb2 in A – the canine ovary (A) and B – the uterus normalised to the mean expression of the Actb and Gapdh housekeeping genes. Bars represent mean ± standard deviation. A, B and C – significant differences between groups (P-value < 0.05). A1 – younger bitches with optimal body weight (body condition score (BCS) = 4/9); A2 – younger confirmed overweight bitches (BCS > 5/9); B1 – elderly bitches with optimal body weight (BCS = 4/9); B2 – elderly confirmed overweight bitches (BCS > 5/9)
Relative expression of Nucb2 in A – the canine ovary (A) and B – the uterus normalised to the mean expression of the Actb and Gapdh housekeeping genes. Bars represent mean ± standard deviation. A, B and C – significant differences between groups (P-value < 0.05). A1 – younger bitches with optimal body weight (body condition score (BCS) = 4/9); A2 – younger confirmed overweight bitches (BCS > 5/9); B1 – elderly bitches with optimal body weight (BCS = 4/9); B2 – elderly confirmed overweight bitches (BCS > 5/9)

Fig. 2.

Concentrations of nesfatin-1 in peripheral blood from healthy bitches in optimal body condition (A1 and B1 groups) and overweight bitches (A2 and B2 groups). Bars represent the mean ± standard deviation. A and B – significant differences between groups (P-value < 0.05). A1 – younger bitches with optimal body weight (BCS = 4/9); A2 – younger confirmed overweight bitches (BCS > 5/9); B1 – elderly bitches with optimal body weight (BCS = 4/9); B2 – elderly confirmed overweight bitches (BCS > 5/9)
Concentrations of nesfatin-1 in peripheral blood from healthy bitches in optimal body condition (A1 and B1 groups) and overweight bitches (A2 and B2 groups). Bars represent the mean ± standard deviation. A and B – significant differences between groups (P-value < 0.05). A1 – younger bitches with optimal body weight (BCS = 4/9); A2 – younger confirmed overweight bitches (BCS > 5/9); B1 – elderly bitches with optimal body weight (BCS = 4/9); B2 – elderly confirmed overweight bitches (BCS > 5/9)

Fig. 3.

A – Representative canine ovary sections showing nesfatin-1 localisation (in red) and cell nuclei stained by 4′,6-diamidino-2-phenylindole (DAPI – in blue). Signal detection of nesfatin-1 in the ovaries of bitches with body condition score (BCS) = 4/9 (a, b and c) and overweight bitches with BCS > 5/9 (d, e and f). Scale bar – 50 μm. NC – negative control; B – Level of nesfatin-1 immunoreactivity analysed in a group of bitches with BCS = 4/9 and overweight bitches with BCS > 5/9. CL – corpus luteum; F – ovarian follicles; SC – ovarian cortical stromal cells. Bars represent the mean level of neuropeptide immunoreactivity ± standard deviation. A, B and C – statistically significant difference between groups at P-value < 0.05. SC-1, F-1 and CL-1 – dogs with BCS = 4/9; SC-2, F-2 and CL-2 – dogs with BCS > 5/9. C – haematoxylin-eosin staining in a representative photomicrograph of an ovarian section from healthy female dogs in the dioestrous phase; arrows – ovarian follicles; asterisk – corpus luteum. Scale bar – 200 μm
A – Representative canine ovary sections showing nesfatin-1 localisation (in red) and cell nuclei stained by 4′,6-diamidino-2-phenylindole (DAPI – in blue). Signal detection of nesfatin-1 in the ovaries of bitches with body condition score (BCS) = 4/9 (a, b and c) and overweight bitches with BCS > 5/9 (d, e and f). Scale bar – 50 μm. NC – negative control; B – Level of nesfatin-1 immunoreactivity analysed in a group of bitches with BCS = 4/9 and overweight bitches with BCS > 5/9. CL – corpus luteum; F – ovarian follicles; SC – ovarian cortical stromal cells. Bars represent the mean level of neuropeptide immunoreactivity ± standard deviation. A, B and C – statistically significant difference between groups at P-value < 0.05. SC-1, F-1 and CL-1 – dogs with BCS = 4/9; SC-2, F-2 and CL-2 – dogs with BCS > 5/9. C – haematoxylin-eosin staining in a representative photomicrograph of an ovarian section from healthy female dogs in the dioestrous phase; arrows – ovarian follicles; asterisk – corpus luteum. Scale bar – 200 μm

Fig. 4.

A – Representative canine uterus sections showing nesfatin-1 localisation (in red) and cell nuclei stained by 4′,6-diamidino-2-phenylindole (DAPI – in blue). Signal detection of nesfatin-1 in the uteri of bitches with body condition score (BCS) = 4/9 (a and b) and overweight individuals with BCS > 5/9 (c and d). Scale bar – 100 μm (a and c), 50 μm (b, d and negative control (NC)), 20 μm (positive control (PC)). B – Level of nesfatin-1 immunoreactivity in the endometrial epithelium (LE), the glandular epithelium (GE), and endometrial stromal cells (SC). Bars represent the mean level of neuropeptide immunoreactivity ± standard deviation. A, B and C – statistically significant difference between groups at P-value < 0.05. LE-1, GE-1 and SC-1 – dogs with BCS = 4/9; LE-2, GE-2 and SC-2 – dogs with BCS > 5/9. C – Representative photomicrograph of histological staining with haematoxylin and eosin of a uterine section from a healthy female dog in the dioestrous phase; M – myometrium; E – endometrium; Scale bar – 200 μm
A – Representative canine uterus sections showing nesfatin-1 localisation (in red) and cell nuclei stained by 4′,6-diamidino-2-phenylindole (DAPI – in blue). Signal detection of nesfatin-1 in the uteri of bitches with body condition score (BCS) = 4/9 (a and b) and overweight individuals with BCS > 5/9 (c and d). Scale bar – 100 μm (a and c), 50 μm (b, d and negative control (NC)), 20 μm (positive control (PC)). B – Level of nesfatin-1 immunoreactivity in the endometrial epithelium (LE), the glandular epithelium (GE), and endometrial stromal cells (SC). Bars represent the mean level of neuropeptide immunoreactivity ± standard deviation. A, B and C – statistically significant difference between groups at P-value < 0.05. LE-1, GE-1 and SC-1 – dogs with BCS = 4/9; LE-2, GE-2 and SC-2 – dogs with BCS > 5/9. C – Representative photomicrograph of histological staining with haematoxylin and eosin of a uterine section from a healthy female dog in the dioestrous phase; M – myometrium; E – endometrium; Scale bar – 200 μm

Primer sequences

Gene namePrimer sequenceGenBank accession No. of source geneBase pairsReference
Nucb2F: 5′-CAAGTGATTGATGTGCTGGAA-3′R: 5′-GCCACTTGTTGTCTTTTCAGTTC-3′XM_534078158(19)
GapdhF: 5′-GGGTCATCATCTCTGCTCCT-3′R: 5′-AGTGGTCATGGATGACTTTGG-3′AF327898.1150(26)
ActbF: 5′-CTGGACTTCGAGCAGGAGAT-3′R: 5′-GATACCGCATGATTCCATCC-3′AF021873.2161(26)

Details of the bitches used in the study

GroupA1A2B1B2
Body condition score4–5/96–8/94–5/96–8/9
Number of bitchesn = 16n = 14n = 16n = 14
Age (years)
       Max.32.812.514.0
       Min.1.21.06.56.3
       Mean ± standard deviation2.06 ± 1.392.2 ± 0.928.3 ± 2.610.73 ± 2.34
Body weight (kg)
       Max.23352543.5
       Min.2.564.34.2
       mean ± standard deviation8.07 ± 6.7316.24 ± 7.0710.5 ± 6.719.7 ± 13.71
Breed
       mixed-breed5444
       purebred11101210
Progesterone (ng/mL)
       Max.25.9336.6539.0349.98
       Min.1.2771.4391.8561.495
       Mean ± standard deviation6.50 ± 4.309.795 ± 8.5411.70 ± 9.6812.44 ± 11.74
DOI: https://doi.org/10.2478/jvetres-2025-0015 | Journal eISSN: 2450-8608
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Language: English
Page range: 141 - 150
Submitted on: Aug 18, 2024
Accepted on: Mar 10, 2025
Published on: Mar 25, 2025
Published by: National Veterinary Research Institute in Pulawy
In partnership with: Paradigm Publishing Services
Publication frequency: 4 times per year
Keywords:
body condition score,
canine,
nesfatin-1,
ovary,
uterus
Related subjects:
Life sciences,
Molecular biology,
Microbiology and virology,
Life sciences, other,
Medicine,
Veterinary medicine

© 2025 Marta Rybska, Marek Skrzypski, Karolina Pusiak, Tatiana Wojciechowicz, Adam Mieldzioc, published by National Veterinary Research Institute in Pulawy
This work is licensed under the Creative Commons Attribution 4.0 License.

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