Perinatal protein malnutrition induces the emergence of enduring effects and age-related impairment behaviors, increasing the death risk in a mouse model

ABSRACT

Background

Early-life adversity impacts on the offspring’s brain development and is associated with a higher risk of developing age-associated diseases. In particular, perinatal protein malnutrition appears to be one of the most critical nutritional deficiencies affecting the individual’s health and survival, but little is known about its effects on the persistence of behavioral alterations throughout life. Thus, the aim of the present study was to investigate how perinatal protein malnutrition impacts on age-related changes in the neuromuscular, cognitive and behavioral functions throughout life in a mouse model.

Methods

One group of CF-1 dams received a normal-protein diet (NP: 20% casein) during gestation and lactation, whereas another group received a low-protein diet (LP: 10% casein). The offspring of both groups were analyzed by means of several behavioral tests at four different ages (young: 6–10 weeks old, mature: 22–26 weeks old, middle age: 39–43 weeks old, and old: 55–59 weeks old).

Results

Regarding neuromuscular functions, LP mice showed an early deterioration in muscular strength and a reduction in the body weight throughout life. Regarding behavior, while NP mice showed an age-related reduction of exploratory behavior, LP mice showed a constantly low level of this behavior, as well as high anxiety-like behavior, which remained at high levels throughout life. Regarding cognitive functions, LP mice showed deteriorated working memory at middle age. Finally, LP mice died 3.4 times earlier than NP mice. Analysis of the sex-related vulnerability showed that females and males were equally affected by perinatal protein malnutrition throughout life.

Conclusion

Our results demonstrate that perinatal protein malnutrition induces enduring and age-related impairment behaviors, which culminate in higher death risk, affecting males and females equally.

KEYWORDS:

• Early-life adversity
• aging
• sex vulnerability
• early-life stress
• anxiety-like behavior
• exploratory behavior
• cognitive skills
• neuromuscular functions

Disclosure statement

No potential conflict of interest was reported by the author(s).

Ethics approval

This study was performed in accordance with local regulations and the National Institutes of Health (NIH) Guide of the Care and Use of Laboratory Animals (NIH publication 80-23/96) and previously approved by the Ethics Committee (CICUAL – Protocol N° 0024 approved on September 13th, 2013) of the Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires.

Additional information

Funding

This work was supported by the Agencia Nacional de Promoción Científica y Tecnológica (ANPCYT) [grant number PICT2013-0653], Universidad de Buenos Aires (UBA) [grant number UBACYT2014-011] and Consejo Nacional de Investigaciones Científicas y Técnicas (AR) (CONICET) [grant number PIP 2012-0476]. Fondo para la Investigación Científica y Tecnológica (AR); Secretaria de Ciencia y Tecnica, Universidad de Buenos Aires.

Notes on contributors

Nadina M. Ferroni

Nadina M. Ferroni: I obtained a Biological Sciences degree with an orientation in animal physiology from the University of Buenos Aires. I have been working in the laboratory of Neuroepigenetics of Dr. Eduardo Cánepa (School of Exact and Natural Sciences, University of Buenos Aires) since 2014 where I did my undergraduated thesis (Universtity of Buenos Aires fellowship) and started my PhD (CONICET fellowship). I have been studying the effect of perinatal protein malnutrition as an early-life adversity that can modify and accelerate the aging process. I have focused on the age-related behavioral deterioration. Nowadays, under Dr. Silvina Sonzogni and Dr. Eduardo Cánepa supervision, I am studying the molecular and cellular mechanisms underlying this process: like increased oxidative stress, accelerated senescence and altered gene expression.

Bruno G. Berardino

Bruno G. Berardino: I am a postdoc researcher (CONICET fellow) studying the effects of early adversities in the development of mental health disorders and the mediation of epigenetic mechanisms in the incorporation of those experiences into the brain. This project arises from previous studies at Dr. Eduardo T. Cánepa’s lab in which we proposed to study the causes of the persistence throughout life of cognitive deficiencies originated in adverse events occurred at early stages of the development in a perinatal malnutrition mouse model. During my PhD (CONICET fellow) I studied the role of microRNAs in the establishment of behavioral effects derived from perinatal malnutrition (Belluscio et al., 2014; Berardino et al., 2017; Berardino et al., 2019). During this period, I obtained a Fulbright scholarship to analyze oligodendrocyte morphology in Gabriel Corfas’ lab at Boston Children’s Hospital (Harvard University). Actually, I successfully administer a research project (staffing and budget), collaborate with other researchers, and produce peer-reviewed publications (Ogara et al., 2014; Fiszbein et al., 2016; Romero et al., 2017; Pregi et al., 2017; Berardino, Fesser et al., 2019). In addition, I am a Teaching Assistant in Molecular Biology (University of Buenos Aires) and participate in community-based activities related to scientific communication.

Laura M. Belluscio

Laura M. Belluscio: I completed my biology degree with an orientation in molecular biology at the University of Buenos Aires. After working in research for two years I switched my focus to Neuroscience. I wanted to get out of the cell and try to understand a bit more about how systems work. During my PhD I worked in the setup of a perinatal protein malnutrition model in mice. The model aimed at unraveling the relationship between behavioral changes observed in malnourished mice and their molecular correlates.

María S. Fernández

María S. Fernández: From 2003 I am a Teaching Assistant from statistics in the Ecology, Genetics and Evolution Department (DEGE) from the University of Buenos Aires (UBA). In 2012 I obtained my PhD and them I became a researcher from the National Research Council (CONICET) where I work in epidemiology, first in neglected diseases and actually in non-communicable diseases.

Estefanía A. Fesser

Estefanía A. Fesser: I am graduated in Genetics at the School of Agrarian, Natural and Environmental Sciences of the University of the Northwest of Buenos Aires (UNNOBA), Argentina. I completed the Final Degree Project in Biotechnology Laboratory of the National Institute of Agricultural Technology (INTA), Pergamino, Buenos Aires. Since 2015, I have been working as a PhD fellow in the Neuroepigenetic laboratory of Dr. Eduardo Cánepa at the School of Sciences, University of Buenos Aires (UBA) (https://www.neuroepigenetica.qb.fcen.uba.ar). My thesis work aims to study the epigenetic mechanisms that mediate the deficiencies in social cognition and its intergenerational transmission derived from perinatal malnutrition in a murine model. In addition, I participate in research lines mainly dedicated to assessing the effects of other early adversities as abusive consumption of cocaine base paste, paternal consumption of alcohol and exposition to social vulnerability. I will have finished my PhD by 2020 and I would like to continue my career doing a post-PhD about additions focused on translational research.

Silvina V. Sonzogni

Silvina V. Sonzogni: I started my research career in the laboratory of Applied Molecular Biology, National University of Misiones, where I obtained the ‘Licenciatura’ Degree in Genetics. I worked on the epidemiological study of cervical human papillomavirus (HPV) infection in women from Misiones (Argentina). In 2007, I started working as a Ph.D. fellow in the laboratory of Dr. Eduardo Cánepa, (School of Exact and Natural Sciences, University of Buenos Aires). My thesis work, was aimed at studying the role of p19INK4d in the induction of genotoxic and replicative senescence. I learned and gained experience in molecular biology techniques and I have worked with mammalian cell cultures. At the end of 2011, I obtained a postdoctoral fellowship awarded from CONICET. I continued as a postdoc in Dr. Cánepa’s laboratory, my work has focused on the DNA damage and chromatin accessibility. Finishing my postdoc, I started to work with a mouse model of perinatal protein malnutrition particularly variations of mouse maternal care associated to malnutrition. Actually, as Assistant Researcher I’m studying the molecular basis of long-term effects of perinatal protein malnutrition. My research interests are particularly directed towards the age-associated changes established in early life.

Eduardo T. Cánepa

Eduardo T. Cánepa: I am Associate Professor and Principal Researcher at and Head of the School of Exact and Natural Sciences of the University of Buenos Aires, and head of the Neuroepigenetic group. My research project is aimed to investigate the molecular causes of the persistence throughout life of cognitive deficiencies derived from protein malnutrition during critical periods of development and maturation of the brain. This issue is of huge importance because it involves the problem of malnutrition, which is present in large parts of the population of our country and affects mainly the poorer sectors of the population. I have extensive training in Molecular Biology and Neurobiology as well as in key research areas to carry out the proposed project.