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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

Marta Rybska

Marta Rybska

Department of Preclinical Sciences and Infectious Diseases, Poznań University of Life SciencesPoznań, Poland
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Rybska, Marta
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Marek Skrzypski

Marek Skrzypski

Department of Animal Physiology, Biochemistry and Biostructure, Poznań University of Life SciencesPoznań, Poland
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Skrzypski, Marek
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Karolina Pusiak

Karolina Pusiak

Department of Preclinical Sciences and Infectious Diseases, Poznań University of Life SciencesPoznań, Poland
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Pusiak, Karolina
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Tatiana Wojciechowicz

Tatiana Wojciechowicz

Department of Animal Physiology, Biochemistry and Biostructure, Poznań University of Life SciencesPoznań, Poland
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Wojciechowicz, Tatiana
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Adam Mieldzioc

Adam Mieldzioc

Department of Mathematical and Statistical Methods, Poznań University of Life SciencesPoznań, Poland
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Mieldzioc, Adam
  
25 mar 2025
The tissue distribution of nucleobindin-2/nesfatin-1 in the reproductive organs of bitches with regard to the animal’s age and body weight's Cover Image
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Publicado en línea: 25 mar 2025
Páginas: 141 - 150
Recibido: 18 ago 2024
Aceptado: 10 mar 2025
DOI: https://doi.org/10.2478/jvetres-2025-0015
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© 2025 Marta Rybska et al., published by Sciendo
This work is licensed under the Creative Commons Attribution 4.0 International License.

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
Idioma:
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Calendario de la edición:
4 veces al año
Temas de la revista:
Ciencias de la vida, Biología molecular, Microbiología y virología, Ciencias de la vida, otros, Medicina, Medicina veterinaria
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