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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

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25 mar 2025

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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 name Primer sequence GenBank accession No. of source gene Base pairs Reference
Nucb2 F: 5′-CAAGTGATTGATGTGCTGGAA-3′R: 5′-GCCACTTGTTGTCTTTTCAGTTC-3′ XM_534078 158 (19)
Gapdh F: 5′-GGGTCATCATCTCTGCTCCT-3′R: 5′-AGTGGTCATGGATGACTTTGG-3′ AF327898.1 150 (26)
Actb F: 5′-CTGGACTTCGAGCAGGAGAT-3′R: 5′-GATACCGCATGATTCCATCC-3′ AF021873.2 161 (26)

Details of the bitches used in the study

Group A1 A2 B1 B2
Body condition score 4–5/9 6–8/9 4–5/9 6–8/9
Number of bitches n = 16 n = 14 n = 16 n = 14
Age (years)
       Max. 3 2.8 12.5 14.0
       Min. 1.2 1.0 6.5 6.3
       Mean ± standard deviation 2.06 ± 1.39 2.2 ± 0.92 8.3 ± 2.6 10.73 ± 2.34
Body weight (kg)
       Max. 23 35 25 43.5
       Min. 2.5 6 4.3 4.2
       mean ± standard deviation 8.07 ± 6.73 16.24 ± 7.07 10.5 ± 6.7 19.7 ± 13.71
Breed
       mixed-breed 5 4 4 4
       purebred 11 10 12 10
Progesterone (ng/mL)
       Max. 25.93 36.65 39.03 49.98
       Min. 1.277 1.439 1.856 1.495
       Mean ± standard deviation 6.50 ± 4.30 9.795 ± 8.54 11.70 ± 9.68 12.44 ± 11.74
Język:
Angielski
Częstotliwość wydawania:
4 razy w roku
Dziedziny czasopisma:
Nauki biologiczne, Biologia molekularna, Mikrobiologia i wirusologia, Nauki biologiczne, inne, Medycyna, Weterynaria