Cervical Lymph Duct-Cannulated Rat Model for Assessing Lymphatic Transport from the Head and Brain

Zihan Zhu; Hefeng Song; Wei Liu; Mingliang Tian; Thu Hoang; Sanjeevini B Reddiar; Natalie L Trevaskis; Sifei Han; Luojuan Hu

doi:10.3791/70217

Method Article

Cervical Lymph Duct-Cannulated Rat Model for Assessing Lymphatic Transport from the Head and Brain

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DOI:

10.3791/70217

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March 10th, 2026

Zihan Zhu¹^,² , Hefeng Song³ , Wei Liu¹ , Mingliang Tian¹ , Thu Hoang³ , Sanjeevini B Reddiar³ , Natalie L Trevaskis³ , Sifei Han¹^,²^,³ , Luojuan Hu¹^,²

¹State Key Laboratory of Natural Medicines, School of Pharmacy, China Pharmaceutical University, ²MOE Joint International Research Laboratory of Target Discovery and New Drug Innovation, China Pharmaceutical University, ³Department of Drug Delivery, Disposition and Dynamics, Monash Institute of Pharmaceutical Sciences, Monash University

Summary

This experimental protocol demonstrates a deep cervical lymph duct-cannulated rat model that enables direct and quantitative assessment of lymphatic drainage from the head region. The model facilitates detailed analysis of central nervous system (CNS)-derived solutes, immune cell populations, and the distribution and clearance of CNS-targeted therapeutics.

Abstract

Lymph flows from the central nervous system (CNS) via a series of lymphatic vessels (LVs) and lymph nodes that converge at the cervical lymph ducts. In addition to maintaining fluid balance, these lymphatic ducts play a key role in transporting a wide array of substances, including endogenous metabolites, signaling molecules, immune cells, and small and macromolecular drugs. This is critical for physiological homeostasis and immune function in both intracranial (e.g., brain) and extracranial regions (e.g., nasal and oral cavities). Impaired lymphatic drainage from the brain has increasingly been linked to a range of neurological and neurodegenerative disorders. Cervical lymph duct cannulation enables the collection of lymph draining the head and brain, allowing the measurement of the concentration and transport rate of various substances via the lymphatic system. Changes in these factors in response to different challenges (e.g., drugs, stress, trauma) and diseases (e.g., stroke, infection, Alzheimer's disease) can also be determined. Here, we describe an anesthetized, deep cervical lymph duct cannulated rat model that enables lymph collection for several hours following surgery. The method may be combined with imaging and multi-omics technologies for the measurement of a wide range of parameters of interest in the lymph. This facilitates fundamental physiological and pathophysiological research of the head and neck region, as well as pharmacokinetic/pharmacodynamic studies of drugs, particularly for the treatment of CNS diseases.

Introduction

The lymphatic system, which is distributed throughout the body, is essential for maintaining fluid homeostasis, regulating immune responses, and facilitating the transport of nutrients and signaling molecules¹. The peripheral lymphatics begin in the lymphatic capillaries, which collect interstitial fluid and solutes and drain into larger collecting lymphatic vessels (LVs) that possess a surrounding smooth muscle layer; these vessels then deliver lymph to local lymph nodes for immune processing²^,³. The lymphatic network draining the head region is important for the homeostasis of both in....