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. 2021 Jan-Feb;12(1):133-148.
doi: 10.32598/bcn.12.1.2346.1. Epub 2021 Jan 1.

ComparativeStudyofCRHMicroinjections Into PVN and CeA Nuclei on Food Intake, Ghrelin, Leptin, and Glucose Levels in Acute Stressed Rats

Maryam Radahmadi  1 , Mina Sadat Izadi  1 , Atefeh Rayatpour  1 , Maedeh Ghasemi  1
Affiliations

ComparativeStudyofCRHMicroinjections Into PVN and CeA Nuclei on Food Intake, Ghrelin, Leptin, and Glucose Levels in Acute Stressed Rats

Maryam Radahmadi et al. Basic Clin Neurosci. 2021 Jan-Feb.

Abstract

Introduction: Corticotropin-Releasing Hormone (CRH) is involved in stress and energy homeostasis. On the other hand, CRH receptors also exist within the paraventricular nucleus (PVN) and Central Amygdala (CeA) nuclei. The present study compared the effect of CRH microinjections into PVN and CeA on three consecutive hours and cumulative food intake, internal regulatory factors of food intake, such as serum leptin and ghrelin, as well as blood glucose levels in rats under different acute psychological (Social Stress [SS] and Isolation Stress [IS] group) stresses.

Methods: Sixty-six male Wistar rats were randomly allocated to 11 groups: Control, Sham, CRH-PVN, CRH-CeA, SS, IS, SS-CRH-PVN, SS-CRH-CeA, IS-CRH-PVN, and IS-CRH-CeA groups. The CRH (2 µg/kg in 0.5 µL saline) was injected into PVN and CeA nuclei in rats under everyday, acute social stress and isolation stress conditions.

Results: Acute isolation and social stresses did not affect cumulative food intake. Whereas isolation stress led to changes in both leptin and glucose levels, social stress reduced only glucose levels. Cumulative food intake significantly decreased under acute CRH injection into the CeA and particularly into the PVN. Blood glucose significantly reduced in all the groups receiving CRH into their CeA.

Conclusion: The PVN played a more important role compared to CeA on food intake. These nuclei probably employ different mechanisms for their effects on food intake. Besides, it seems that exogenously CRH injection into the PVN probably had a more anorectic effect than naturally activated CRH by stresses. Acute isolation stress had a greater impact than social stress on leptin level and cumulative food intake. Thus, elevated food intake related to leptin compared to ghrelin and glucose levels in the CRH-PVN group under acute social stress.

Keywords: Central amygdala; Corticotropin-releasing hormone; Food intake; Isolation stress; Paraventricular; Social stress.

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Conflict of interest statement

Conflict of interest The authors declared no conflict of interest.

Figures

Figure 1.
Figure 1.
Experimental schedule for all groups, stereotaxic surgeries, and the day on which behavioral tests and bleeding were tested, animals exposed to isolation and or social stresses
Figure 2.
Figure 2.
The position of the injection cannula tips A: The Paraventricular Nucleus (PVN); and B: The Central Amygdala (CeA) regions for all rats included in the data analyses.
Figure 3.
Figure 3.
Comparison of the experimental groups in terms of food intake (g) after 1 h Results are expressed as Means±SEM (ANOVA followed by LSD post-hoc test). *P<0.05; and **P<0.01 compared to Co; #P<0.05 compared to Sh-PVN; €€P<0.01 compared to SS; θθP<0.01 compared to IS; ΔP<0.05 compared to CRH-PVN; ӾӾӾP<0.001 compared to CRH-CeA; ǂǂP<0.01 compared to SS-CRH-PVN; ҰҰҰP<0.001 compared to IS-CRH-PVN; £££P<0.001 compared to SS-CRH-CeA. PVN: Paraventricular nucleus; CeA: Central amygdala; Co: Control group; Sh-PVN: Sham-operated PVN group; Sh-CeA: Shamoperated CeA group; SS: Social stress group; IS: Isolation stress group; CRH-PVN: CRH injection into the PVN; CRH-CeA: CRH injection into the CeA; SS-CRH-PVN: Social stress-CRH injection into the PVN; IS-CRH-PVN: Isolation stress-CRH injection into the PVN; SS-CRH-CeA: Social stress-CRH injection into the CeA; ISCRH-CeA: Isolation stress-CRH injection into the CeA.
Figure 4.
Figure 4.
Comparison of the experimental groups in terms of their food intake (g) after 2 h Results are expressed as Means±Standard Error of The Mean (ANOVA followed by LSD post-hoc test). *P<0.05; and ***P<0.001 compared to Co; ##P<0.01 compared to Sh-PVN; €P<0.05; €€P<0.01; and €€€P<0.001compared to SS; θP<0.05 compared to IS; ΔΔP<0.01; and ΔΔΔP<0.001 compared to CRH-PVN; ǂP<0.05 compared to SS-CRH-PVN; £P<0.05 compared to SS-CRH-CeA. PVN: Paraventricular nucleus; CeA: Central amygdala; Co: Control group; Sh-PVN: Sham-operated PVN group; Sh-CeA: Sham-operated CeA group; SS: Social stress group; IS: Isolation stress group; CRH-PVN: CRH injection into the PVN; CRH-CeA: CRH injection into the CeA; SS-CRH-PVN: Social stress-CRH injection into the PVN; IS-CRH-PVN: Isolation stress-CRH injection into the PVN; SS-CRH-CeA: Social stress-CRH injection into the CeA; IS-CRH-CeA: Isolation stress-CRH injection into the CeA.
Figure 6.
Figure 6.
Comparison of the experimental groups in terms of cumulative food intake (g) Results are expressed as Means±Standard Error of the Mean (ANOVA followed by LSD post-hoc test). *P<0.05; and **P<0.01 compared to Co; ##P<0.01 compared to Sh-PVN; ₸P<0.05 compared to Sh-CeA; €P<0.05 compared to SS; θP<0.05 compared to IS; ΔP<0.05 compared to CRH-PVN; ӾӾӾP<0.001 compared to CRH-CeA; ҰҰP<0.01 compared to IS-CRHPVN; £P<0.05 compared to SS-CRH-CeA. PVN: Paraventricular nucleus; CeA: Central amygdala; Co: Control group; Sh-PVN: Sham-operated PVN group; Sh-CeA: Shamoperated CeA group; SS: Social stress group; IS: Isolation stress group; CRH-PVN: CRH injection into the PVN; CRH-CeA: CRH injection into the CeA; SS-CRH-PVN: Social stress-CRH injection into the PVN; IS-CRH-PVN: Isolation stress-CRH injection into the PVN; SS-CRH-CeA: Social stress-CRH injection into the CeA; ISCRH-CeA: Isolation stress-CRH injection into the CeA.
Figure 5.
Figure 5.
Comparison of the experimental groups in terms of food intake (g) after 3 h Results are expressed as Means±SEMean (ANOVA followed by LSD post-hoc test). *P<0.05; **P<0.01; and ***P<0.001 compared to Co; #P<0.05; and ##P<0.01 compared to Sh-PVN; €P<0.05; and €€P<0.01 compared to SS; θP<0.05; and θθθP<0.001 compared to IS; ӾP<0.05; and ӾӾP<0.01 compared to CRH-CeA. PVN: Paraventricular nucleus; CeA; Central amygdala; Co: Control group; Sh-PVN: Sham-operated PVN group; Sh-CeA: Shamoperated CeA group; SS: Social stress group; IS: Isolation stress group; CRH-PVN: CRH injection into the PVN; CRH-CeA: CRH injection into the CeA; SS-CRH-PVN: Social stress-CRH injection into the PVN; IS-CRH-PVN: Isolation stress-CRH injection into the PVN; SS-CRH-CeA: Social stress-CRH injection into the CeA; ISCRH-CeA: Isolation stress-CRH injection into the CeA.
Figure 7.
Figure 7.
Comparison of the experimental groups in terms of serum leptin level (ng/mL) Results are expressed as Means±Standard Error of the Mean (ANOVA followed by LSD post-hoc test). *P<0.05; and **P<0.01 compared to Co; #P<0.05 compared to Sh-PVN; ₸P<0.05 compared to Sh-CeA; €P<0.05; and €€P<0.01 compared to SS; ΔP<0.05 compared to CRH-PVN; £P<0.05 compared to SS-CRH-CeA. PVN: Paraventricular nucleus; CeA: Central amygdala; Co: Control group; Sh-PVN: Sham-operated PVN group; Sh-CeA: Shamoperated CeA group; SS: Social stress group; IS: Isolation stress group; CRH-PVN: CRH injection into the PVN; CRH-CeA: CRH injection into the CeA; SS-CRH-PVN: Social stress-CRH injection into the PVN; IS-CRH-PVN: Isolation stress-CRH injection into the PVN; SS-CRH-CeA: Social stress-CRH injection into the CeA; ISCRH-CeA: Isolation stress-CRH injection into the CeA.
Figure 8.
Figure 8.
Comparison of the experimental groups in terms of serum ghrelin level (ng/mL) Results are expressed as Means±Standard Error of the Mean (ANOVA followed by LSD post-hoc test). PVN: Paraventricular nucleus; CeA: Central amygdala; Co: Control group; Sh-PVN: Sham-operated PVN group; Sh-CeA: Sham-operated CeA group; SS: Social stress group; IS: Isolation stress group; CRH-PVN: CRH injection into the PVN; CRHCeA: CRH injection into the CeA; SS-CRH-PVN: Social stress-CRH injection into the PVN; IS-CRH-PVN: Isolation stress-CRH injection into the PVN; SS-CRH-CeA: Social stress-CRH injection into the CeA; ISCRH-CeA:Isolation stress-CRH injection into the CeA.
Figure 9.
Figure 9.
Comparison of the experimental groups in terms of blood glucose level (mg/dL) Results are expressed as Means±Standard Error of the Mean (ANOVA followed by LSD post-hoc test). *P<0.05; and **P<0.01 compared to Co; ₸P<0.05 compared to Sh-CeA. PVN: Paraventricular nucleus; CeA: Central amygdala; Co: Control group; Sh-PVN: Sham-operated PVN group; Sh-CeA: Shamoperated CeA group; SS: Social stress group; IS: Isolation stress group; CRH-PVN: CRH injection into the PVN; CRH-CeA: CRH injection into the CeA; SS-CRH-PVN: Social stress-CRH injection into the PVN; IS-CRH-PVN: Isolation stress-CRH injection into the PVN; SS-CRH-CeA: Social stress-CRH injection into the CeA; IS-CRH-CeA: Isolation stress-CRH injection into the CeA.
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