Perilipin 5 links mitochondrial uncoupled respiration in brown fat to healthy white fat remodeling and systemic glucose tolerance

Exposure of mice or humans to cold promotes significant changes in brown adipose tissue (BAT) with respect to histology, lipid content, gene expression, and mitochondrial mass and function. Herein we report that the lipid droplet coat protein Perilipin 5 (PLIN5) increases markedly in BAT during exposure of mice to cold. To understand the functional significance of cold-induced PLIN5, we created and characterized gain- and loss-of-function mouse models. Enforcing PLIN5 expression in mouse BAT mimics the effects of cold with respect to mitochondrial cristae packing and uncoupled substrate-driven respiration. PLIN5 is necessary for the maintenance of mitochondrial cristae structure and respiratory function during cold stress. We further show that promoting PLIN5 function in BAT is associated with healthy remodeling of subcutaneous white adipose tissue and improvements in systemic glucose tolerance and diet-induced hepatic steatosis. These observations will inform future strategies that seek to exploit thermogenic adipose tissue as a therapeutic target for type 2 diabetes, obesity, and nonalcoholic fatty liver disease.

The exact sample size (n) for each experimental group/condition, given as a discrete number and unit of measurement A statement on whether measurements were taken from distinct samples or whether the same sample was measured repeatedly The statistical test(s) used AND whether they are one-or two-sided Only common tests should be described solely by name; describe more complex techniques in the Methods section.
A description of all covariates tested A description of any assumptions or corrections, such as tests of normality and adjustment for multiple comparisons A full description of the statistical parameters including central tendency (e.g. means) or other basic estimates (e.g. regression coefficient) AND variation (e.g. standard deviation) or associated estimates of uncertainty (e.g. confidence intervals) For null hypothesis testing, the test statistic (e.g. F, t, r) with confidence intervals, effect sizes, degrees of freedom and P value noted Give P values as exact values whenever suitable.

For Bayesian analysis, information on the choice of priors and Markov chain Monte Carlo settings
For hierarchical and complex designs, identification of the appropriate level for tests and full reporting of outcomes Estimates of effect sizes (e.g. Cohen's d, Pearson's r), indicating how they were calculated Our web collection on statistics for biologists contains articles on many of the points above.

Software and code
Policy information about availability of computer code Data collection

Data analysis
For manuscripts utilizing custom algorithms or software that are central to the research but not yet described in published literature, software must be made available to editors/reviewers. We strongly encourage code deposition in a community repository (e.g. GitHub). See the Nature Research guidelines for submitting code & software for further information.

Data
Policy information about availability of data All manuscripts must include a data availability statement. This statement should provide the following information, where applicable: According to vendor anti guinea pig Perilipin 5 (Progen) is validated to use in western blot and has been used in multiple publications (https://www.progen.com/ProductLeaflet/file/getpdf/name/anti-Perilipin_5_%28C-terminus% 29_guinea_pig_polyclonal%2C_serum.pdf?fileId=454). Additionally we had tested this antibody using Perilipin 5 KO mouse and a specific band in the right size can be found in the control mice but not in the KO mice. anti rabbit Perilipin-5 (Bickel Lab) detailed information regarding validation and images can be found in Methods reference 1, additionally we have tested this antibody using Perilipin 5 KO mice and a specific band in the right size can be found in the control mice but not in the KO mice. According to vendor anti-Ucp1 (Abcam) has been validated to use in western blot (https://www.abcam.com/ucp1-antibody-epr20381-ab209483.html) and has been widely use in multiple publications. Additionally we have tested this antibody using the UCP1 KO mice and a specific band can be found in the control mice and not in the KO mice. anti-GDI was generated by Perry Bickel and detailed information can be found in Methods reference 10. According to vendor (Santa Cruz Biotechnology) anti-Actin has been validated to use in western blot and has been widely use in multiple publications (https://www.scbt.com/p/beta-actin-antibody-c4), anti-total AKT from Santa Cruz Biotechnology is currently unavailable but according to the vendor has been used in multiple publications (https://www.scbt.com/p/akt1-2-antibody-n-19?requestFrom=search). anti-phosphoAKT was purchased from During the execution of some experiments and when possible, investigators were blinded to genotype of the mice. For histology and electron microscopy, the investigator was blinded to mouse genotype during image acquisition. For experiments with chow or high fat diet, blinding was not possible due to obvious differences between diets. In some experiments, blinding was not possible, because the setup and execution of the experiment were performed by the same person.
Mice in the same cohort with different genotypes were randomly allocated to the indicated treatment (e.g. Chow vs HFD) For reproducibility we performed independent experiments at least 2 times (two different cohorts of mice). Some experiments were independently reproduced more than 2 times. All the attemps of replication were succesful.