diff --git a/django/contrib/contenttypes/prefetch.py b/django/contrib/contenttypes/prefetch.py
index b02ed3bae573..97ac60295bb4 100644
--- a/django/contrib/contenttypes/prefetch.py
+++ b/django/contrib/contenttypes/prefetch.py
@@ -3,7 +3,7 @@
 
 
 class GenericPrefetch(Prefetch):
-    def __init__(self, lookup, querysets=None, to_attr=None):
+    def __init__(self, lookup, querysets, to_attr=None):
         for queryset in querysets:
             if queryset is not None and (
                 isinstance(queryset, RawQuerySet)
diff --git a/docs/ref/contrib/contenttypes.txt b/docs/ref/contrib/contenttypes.txt
index fa446796594e..5aad2a4ec726 100644
--- a/docs/ref/contrib/contenttypes.txt
+++ b/docs/ref/contrib/contenttypes.txt
@@ -603,7 +603,7 @@ information.
 
 .. versionadded:: 5.0
 
-.. class:: GenericPrefetch(lookup, querysets=None, to_attr=None)
+.. class:: GenericPrefetch(lookup, querysets, to_attr=None)
 
 This lookup is similar to ``Prefetch()`` and it should only be used on
 ``GenericForeignKey``. The ``querysets`` argument accepts a list of querysets,
diff --git a/docs/releases/5.0.5.txt b/docs/releases/5.0.5.txt
index bb1d732167ba..7e7963e707da 100644
--- a/docs/releases/5.0.5.txt
+++ b/docs/releases/5.0.5.txt
@@ -30,3 +30,6 @@ Bugfixes
 
 * Fixed a bug in Django 5.0 that caused a migration crash when altering a
   ``GeneratedField`` referencing a renamed field (:ticket:`35422`).
+
+* Fixed a bug in Django 5.0 where the ``querysets`` argument of
+  ``GenericPrefetch`` was not required (:ticket:`35426`).
diff --git a/tests/contenttypes_tests/test_models.py b/tests/contenttypes_tests/test_models.py
index 88f715ceffcf..799f1cc58c74 100644
--- a/tests/contenttypes_tests/test_models.py
+++ b/tests/contenttypes_tests/test_models.py
@@ -331,6 +331,14 @@ def test_multidb(self):
 
 
 class GenericPrefetchTests(TestCase):
+    def test_querysets_required(self):
+        msg = (
+            "GenericPrefetch.__init__() missing 1 required "
+            "positional argument: 'querysets'"
+        )
+        with self.assertRaisesMessage(TypeError, msg):
+            GenericPrefetch("question")
+
     def test_values_queryset(self):
         msg = "Prefetch querysets cannot use raw(), values(), and values_list()."
         with self.assertRaisesMessage(ValueError, msg):